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The small molecule indirubin-3'-oxime activates Wnt/β-catenin signaling and inhibits adipocyte differentiation and obesity.

Choi OM, Cho YH, Choi S, Lee SH, Seo SH, Kim HY, Han G, Min DS, Park T, Choi KY - Int J Obes (Lond) (2013)

Bottom Line: We investigated the effect of indirubin-3'-oxime (I3O), which was screened as an activator of the Wnt/β-catenin signaling, on inhibiting the preadipocyte differentiation in vitro and in vivo. 3T3L1 preadipocytes were differentiated with 0, 4 or 20 μM of I3O.The I3O effect on adipocyte differentiation was observed by Oil-red-O staining.I3O inhibited the differentiation of 3T3-L1 cells into mature adipocytes and decreased the expression of adipocyte markers, CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor γ, at both mRNA and protein levels.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea [2] Translational Research Center for Protein Function Control, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea.

ABSTRACT

Objectives: Activation of the Wnt/β-catenin signaling pathway inhibits adipogenesis by maintaining preadipocytes in an undifferentiated state. We investigated the effect of indirubin-3'-oxime (I3O), which was screened as an activator of the Wnt/β-catenin signaling, on inhibiting the preadipocyte differentiation in vitro and in vivo.

Methods: 3T3L1 preadipocytes were differentiated with 0, 4 or 20 μM of I3O. The I3O effect on adipocyte differentiation was observed by Oil-red-O staining. Activation of Wnt/β-catenin signaling in I3O-treated 3T3L1 cells was shown using immunocytochemical and immunoblotting analyses for β-catenin. The regulation of adipogenic markers was analyzed via real-time reverse transcription-PCR (RT-PCR) and immunoblotting analyses. For the in vivo study, mice were divided into five different dietary groups: chow diet, high-fat diet (HFD), HFD supplemented with I3O at 5, 25 and 100 mg kg(-1). After 8 weeks, adipose and liver tissues were excised from the mice and subject to morphometry, real-time RT-PCR, immunoblotting and histological or immunohistochemical analyses. In addition, adipokine and insulin concentrations in serum of the mice were accessed by enzyme-linked immunosorbent assay.

Results: Using a cell-based approach to screen a library of pharmacologically active small molecules, we identified I3O as a Wnt/β-catenin pathway activator. I3O inhibited the differentiation of 3T3-L1 cells into mature adipocytes and decreased the expression of adipocyte markers, CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor γ, at both mRNA and protein levels. In vivo, I3O inhibited the development of obesity in HFD-fed mice by attenuating HFD-induced body weight gain and visceral fat accumulation without showing any significant toxicity. Factors associated with metabolic disorders such as hyperlipidemia and hyperglycemia were also improved by treatment of I3O.

Conclusion: Activation of the Wnt/β-catenin signaling pathway can be used as a therapeutic strategy for the treatment of obesity and metabolic syndrome and implicates I3O as a candidate anti-obesity agent.

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Related in: MedlinePlus

Effects of I3O on the liver of HFD-induced mice. Plasma levels of triglyceride (a), total cholesterol (b), and free fatty acid (c) in the liver of mice after 8 weeks. (d) Immunohistochemical analysis of markers in liver. Values are expressed as means±s.e.m. (n=10). *P<0.05, **P<0.01, ***P<0.001 (Kruskal-Wallis test).
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fig4: Effects of I3O on the liver of HFD-induced mice. Plasma levels of triglyceride (a), total cholesterol (b), and free fatty acid (c) in the liver of mice after 8 weeks. (d) Immunohistochemical analysis of markers in liver. Values are expressed as means±s.e.m. (n=10). *P<0.05, **P<0.01, ***P<0.001 (Kruskal-Wallis test).

Mentions: To further confirm the reduction of hyperlipidemia and hyperglycemia after treatment of I3O, we measured TG levels, TC and FFA levels in the liver. The liver has a vital role in fatty acid synthesis and metabolism. Fatty acid synthesized by the liver is converted to TG and transported to the blood.27 Both TG and TC levels in the liver were significantly reduced following I3O treatment, correlating with the reduction in plasma TG and TC levels (Figures 4a and b). Lastly, FFA levels in the liver were reduced by 18.1, 44.7 and 66.6% in 5, 25 and 100 mg kg−1 mice, respectively (Figure 4c). Overall, I3O inhibited lipogenesis in the liver, thereby decreasing the secretion and mobilization of lipids into the bloodstream.


The small molecule indirubin-3'-oxime activates Wnt/β-catenin signaling and inhibits adipocyte differentiation and obesity.

Choi OM, Cho YH, Choi S, Lee SH, Seo SH, Kim HY, Han G, Min DS, Park T, Choi KY - Int J Obes (Lond) (2013)

Effects of I3O on the liver of HFD-induced mice. Plasma levels of triglyceride (a), total cholesterol (b), and free fatty acid (c) in the liver of mice after 8 weeks. (d) Immunohistochemical analysis of markers in liver. Values are expressed as means±s.e.m. (n=10). *P<0.05, **P<0.01, ***P<0.001 (Kruskal-Wallis test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4125748&req=5

fig4: Effects of I3O on the liver of HFD-induced mice. Plasma levels of triglyceride (a), total cholesterol (b), and free fatty acid (c) in the liver of mice after 8 weeks. (d) Immunohistochemical analysis of markers in liver. Values are expressed as means±s.e.m. (n=10). *P<0.05, **P<0.01, ***P<0.001 (Kruskal-Wallis test).
Mentions: To further confirm the reduction of hyperlipidemia and hyperglycemia after treatment of I3O, we measured TG levels, TC and FFA levels in the liver. The liver has a vital role in fatty acid synthesis and metabolism. Fatty acid synthesized by the liver is converted to TG and transported to the blood.27 Both TG and TC levels in the liver were significantly reduced following I3O treatment, correlating with the reduction in plasma TG and TC levels (Figures 4a and b). Lastly, FFA levels in the liver were reduced by 18.1, 44.7 and 66.6% in 5, 25 and 100 mg kg−1 mice, respectively (Figure 4c). Overall, I3O inhibited lipogenesis in the liver, thereby decreasing the secretion and mobilization of lipids into the bloodstream.

Bottom Line: We investigated the effect of indirubin-3'-oxime (I3O), which was screened as an activator of the Wnt/β-catenin signaling, on inhibiting the preadipocyte differentiation in vitro and in vivo. 3T3L1 preadipocytes were differentiated with 0, 4 or 20 μM of I3O.The I3O effect on adipocyte differentiation was observed by Oil-red-O staining.I3O inhibited the differentiation of 3T3-L1 cells into mature adipocytes and decreased the expression of adipocyte markers, CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor γ, at both mRNA and protein levels.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea [2] Translational Research Center for Protein Function Control, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea.

ABSTRACT

Objectives: Activation of the Wnt/β-catenin signaling pathway inhibits adipogenesis by maintaining preadipocytes in an undifferentiated state. We investigated the effect of indirubin-3'-oxime (I3O), which was screened as an activator of the Wnt/β-catenin signaling, on inhibiting the preadipocyte differentiation in vitro and in vivo.

Methods: 3T3L1 preadipocytes were differentiated with 0, 4 or 20 μM of I3O. The I3O effect on adipocyte differentiation was observed by Oil-red-O staining. Activation of Wnt/β-catenin signaling in I3O-treated 3T3L1 cells was shown using immunocytochemical and immunoblotting analyses for β-catenin. The regulation of adipogenic markers was analyzed via real-time reverse transcription-PCR (RT-PCR) and immunoblotting analyses. For the in vivo study, mice were divided into five different dietary groups: chow diet, high-fat diet (HFD), HFD supplemented with I3O at 5, 25 and 100 mg kg(-1). After 8 weeks, adipose and liver tissues were excised from the mice and subject to morphometry, real-time RT-PCR, immunoblotting and histological or immunohistochemical analyses. In addition, adipokine and insulin concentrations in serum of the mice were accessed by enzyme-linked immunosorbent assay.

Results: Using a cell-based approach to screen a library of pharmacologically active small molecules, we identified I3O as a Wnt/β-catenin pathway activator. I3O inhibited the differentiation of 3T3-L1 cells into mature adipocytes and decreased the expression of adipocyte markers, CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor γ, at both mRNA and protein levels. In vivo, I3O inhibited the development of obesity in HFD-fed mice by attenuating HFD-induced body weight gain and visceral fat accumulation without showing any significant toxicity. Factors associated with metabolic disorders such as hyperlipidemia and hyperglycemia were also improved by treatment of I3O.

Conclusion: Activation of the Wnt/β-catenin signaling pathway can be used as a therapeutic strategy for the treatment of obesity and metabolic syndrome and implicates I3O as a candidate anti-obesity agent.

Show MeSH
Related in: MedlinePlus